Harigaya T, Smith W C, Talamantes F
Biology Board of Studies, Thimann Laboratories, University of California, Santa Cruz 95064.
Endocrinology. 1988 Apr;122(4):1366-72. doi: 10.1210/endo-122-4-1366.
The binding sites for mouse placental lactogen-II (mPL-II) in virgin and pregnant mouse hepatic membranes were analyzed by Scatchard analysis and affinity cross-linking. Competitive binding studies showed that mPL-II and mouse PRL (mPRL) bound to the same receptors in all liver membrane preparations, although the affinity of mPRL binding was lower than that of mPL-II binding. Two classes of receptors for mPL-II, high and low affinity, were found by Scatchard analysis. The concentration of both types of sites in liver membranes increased during pregnancy. In contrast, the affinity of both sites for mPL-II was highest in virgin female mice and declined during pregnancy. Cross-linking of [125I]iodo-mPL-II to maternal liver membranes resulted in the specific labeling of one major protein species of 67,000 daltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Under nonreducing conditions, two bands of approximately 63,000 and 60,000 daltons were apparent. Subtraction of the mol wt of mPL-II (22,000, reduced; 20,000, nonreduced) from the mol wt of the cross-linked complex indicated that the mol wt of the receptor was 45,000 under reducing conditions and 43,000 and 40,000 under nonreducing conditions. These observations suggest that mPL-II receptors may be present in mouse liver membranes in at least two forms.
通过Scatchard分析和亲和交联法,对未孕和怀孕小鼠肝细胞膜中鼠胎盘催乳素-II(mPL-II)的结合位点进行了分析。竞争性结合研究表明,在所有肝细胞膜制剂中,mPL-II和鼠催乳素(mPRL)都与相同的受体结合,尽管mPRL结合的亲和力低于mPL-II结合的亲和力。通过Scatchard分析发现了两类mPL-II受体,即高亲和力和低亲和力受体。肝细胞膜中这两种类型位点的浓度在怀孕期间增加。相反,这两种位点对mPL-II的亲和力在未孕雌性小鼠中最高,在怀孕期间下降。在还原条件下,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定,[125I]碘代-mPL-II与母体肝细胞膜的交联导致一种主要的67,000道尔顿蛋白质的特异性标记。在非还原条件下,两条约63,000和60,000道尔顿的条带明显可见。从交联复合物的分子量中减去mPL-II的分子量(还原时为22,000;非还原时为20,000)表明,在还原条件下受体的分子量为45,000,在非还原条件下为43,000和40,000。这些观察结果表明,mPL-II受体可能至少以两种形式存在于小鼠肝细胞膜中。